High ‐speed volumetric imaging in vivo based on structured illumination microscopy with interleaved reconstruction

2021 ◽  
Author(s):  
Ruheng Shi ◽  
Yuting Li ◽  
Lingjie Kong
Keyword(s):  
High Speed ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Volumetric Imaging

scholarly journals HiLo Based Line Scanning Temporal Focusing Microscopy for High-Speed, Deep Tissue Imaging

Membranes ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 634
Author(s):  
Ruheng Shi ◽  
Yuanlong Zhang ◽  
Tiankuang Zhou ◽  
Lingjie Kong
Keyword(s):  
High Speed ◽  
Three Dimensions ◽  
Tissue Imaging ◽  
Deep Penetration ◽  
High Temporal Resolution ◽  
Structured Illumination ◽  
Volumetric Imaging ◽  
Temporal Focusing ◽  
Line Scanning

High-speed, optical-sectioning imaging is highly desired in biomedical studies, as most bio-structures and bio-dynamics are in three-dimensions. Compared to point-scanning techniques, line scanning temporal focusing microscopy (LSTFM) is a promising method that can achieve high temporal resolution while maintaining a deep penetration depth. However, the contrast and axial confinement would still be deteriorated in scattering tissue imaging. Here, we propose a HiLo-based LSTFM, utilizing structured illumination to inhibit the fluorescence background and, thus, enhance the image contrast and axial confinement in deep imaging. We demonstrate the superiority of our method by performing volumetric imaging of neurons and dynamical imaging of microglia in mouse brains in vivo.

High-Speed Structured Illumination Microscopy and Its Applications

2020 ◽  
Vol 57 (24) ◽  
pp. 240001
Author(s):  
赵天宇 Zhao Tianyu ◽  
汪召军 Wang Zhaojun ◽  
冯坤 Feng Kun ◽  
梁言生 Liang Yansheng ◽  
何旻儒 He Minru ◽  
...  
Keyword(s):  
High Speed ◽  
Structured Illumination ◽  
Structured Illumination Microscopy

In-vivo volumetric imaging of the cellular structure of healthy and pathological human cornea with high-speed UHR-OCT (Conference Presentation)

2018 ◽  
Author(s):  
Zohreh Hosseinaee ◽  
Bingyao Tan ◽  
Kirsten Carter ◽  
Denise Hileeto ◽  
Luigina Sorbara ◽  
...  
Keyword(s):  
Cellular Structure ◽  
High Speed ◽  
Human Cornea ◽  
Volumetric Imaging

scholarly journals High-speed multiplane structured illumination microscopy of living cells using an image-splitting prism

Nanophotonics ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 143-148
Author(s):  
Adrien Descloux ◽  
Marcel Müller ◽  
Vytautas Navikas ◽  
Andreas Markwirth ◽  
Robin van den Eynde ◽  
...  
Keyword(s):  
High Speed ◽  
Three Dimensional ◽  
Super Resolution ◽  
Optical Element ◽  
Spatial Light Modulators ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Image Stack ◽  
Light Modulators ◽  
3D Information

AbstractSuper-resolution structured illumination microscopy (SR-SIM) can be conducted at video-rate acquisition speeds when combined with high-speed spatial light modulators and sCMOS cameras, rendering it particularly suitable for live-cell imaging. If, however, three-dimensional (3D) information is desired, the sequential acquisition of vertical image stacks employed by current setups significantly slows down the acquisition process. In this work, we present a multiplane approach to SR-SIM that overcomes this slowdown via the simultaneous acquisition of multiple object planes, employing a recently introduced multiplane image splitting prism combined with high-speed SIM illumination. This strategy requires only the introduction of a single optical element and the addition of a second camera to acquire a laterally highly resolved 3D image stack. We demonstrate the performance of multiplane SIM by applying this instrument to imaging the dynamics of mitochondria in living COS-7 cells.

scholarly journals Live-cell superresolution microscopy reveals the organization of RNA polymerase in the bacterial nucleoid

2015 ◽  
Vol 112 (32) ◽  
pp. E4390-E4399 ◽  
Author(s):  
Mathew Stracy ◽  
Christian Lesterlin ◽  
Federico Garza de Leon ◽  
Stephan Uphoff ◽  
Pawel Zawadzki ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Single Molecule ◽  
Spatial Organization ◽  
Search Time ◽  
Population Level ◽  
Bacterial Cells ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Superresolution Microscopy

Despite the fundamental importance of transcription, a comprehensive analysis of RNA polymerase (RNAP) behavior and its role in the nucleoid organization in vivo is lacking. Here, we used superresolution microscopy to study the localization and dynamics of the transcription machinery and DNA in live bacterial cells, at both the single-molecule and the population level. We used photoactivated single-molecule tracking to discriminate between mobile RNAPs and RNAPs specifically bound to DNA, either on promoters or transcribed genes. Mobile RNAPs can explore the whole nucleoid while searching for promoters, and spend 85% of their search time in nonspecific interactions with DNA. On the other hand, the distribution of specifically bound RNAPs shows that low levels of transcription can occur throughout the nucleoid. Further, clustering analysis and 3D structured illumination microscopy (SIM) show that dense clusters of transcribing RNAPs form almost exclusively at the nucleoid periphery. Treatment with rifampicin shows that active transcription is necessary for maintaining this spatial organization. In faster growth conditions, the fraction of transcribing RNAPs increases, as well as their clustering. Under these conditions, we observed dramatic phase separation between the densest clusters of RNAPs and the densest regions of the nucleoid. These findings show that transcription can cause spatial reorganization of the nucleoid, with movement of gene loci out of the bulk of DNA as levels of transcription increase. This work provides a global view of the organization of RNA polymerase and transcription in living cells.

scholarly journals A Multifocal Multiphoton Volumetric Imaging Technique for High Speed Time-Resolved FRET Imaging in vivo

2016 ◽  
Vol 110 (3) ◽  
pp. 165a ◽  
Author(s):  
Simon P. Poland ◽  
James A. Levitt ◽  
Nikola Krstajić ◽  
Ahmet Erdogen ◽  
Richard J. Walker ◽  
...  
Keyword(s):  
High Speed ◽  
Imaging Technique ◽  
Time Resolved ◽  
Volumetric Imaging

scholarly journals Simulating digital micromirror devices for patterning coherent excitation light in structured illumination microscopy

2020 ◽  
Author(s):  
Mario Lachetta ◽  
Hauke Sandmeyer ◽  
Alice Sandmeyer ◽  
Jan Schulte am Esch ◽  
Thomas Huser ◽  
...  
Keyword(s):  
High Speed ◽  
Consumer Electronics ◽  
Light Sources ◽  
Structured Illumination ◽  
Light Modulation ◽  
Coherent Light ◽  
Structured Illumination Microscopy ◽  
Excitation Light ◽  
Coherent Excitation ◽  
Digital Micromirror Devices

SummaryDigital micromirror devices (DMDs) are spatial light modulators that employ the electro-mechanical movement of miniaturized mirrors to steer and thus modulate the light reflected of a mirror array. Their wide availability, low cost and high speed make them a popular choice both in consumer electronics such as video projectors, and scientific applications such as microscopy.High-end fluorescence microscopy systems typically employ laser light sources, which by their nature provide coherent excitation light. In super-resolution microscopy applications that use light modulation, most notably structured illumination microscopy (SIM), the coherent nature of the excitation light becomes a requirement to achieve optimal interference pattern contrast. The universal combination of DMDs and coherent light sources, especially when working with multiple different wavelengths, is unfortunately not straight forward. The substructure of the tilted micromirror array gives rise to a blazed grating, which has to be understood and which must be taken into account when designing a DMD-based illumination system.Here, we present a set of simulation frameworks that explore the use of DMDs in conjunction with coherent light sources, motivated by their application in SIM, but which are generalizable to other light patterning applications. This framework provides all the tools to explore and compute DMD-based diffraction effects and to simulate possible system alignment configurations computationally, which simplifies the system design process and provides guidance for setting up DMD-based microscopes.

Sign in / Sign up

Export Citation Format

Share Document